Fig. 6

Fowlerstefin induces TNF, IL-1α, IL-1β and IL-6 expression in BV-2 cells. a RT-PCR analysis of TNF, IL-1α, IL-1β and IL-6 mRNA expression in BV-2 cells. BV-2 cells were treated with fowlerstefin (15 µg/ml) and harvested after 0–12 h. Total RNA was isolated from the cells and the expression of TNF, IL-1α, IL-1β and IL-6 was assessed by RT-PCR. GAPDH was used as an internal control. Lane NC: control not treated with fowlerstefin; Lane LPS: LPS (3 µg/ml); Lanes 3, 6, 9 and 12: treated with fowlerstefin for the indicated times. LPS was treated with the cells for 2 h. A representative gel image of three independent experiments which revealed similar patterns is presented. b Quantitative analysis of TNF, IL-1α, IL-1β and IL-6 relative to GAPDH. Graphs show the mean ± SD densitometric ratios of cytokines to GAPDH of three independent experiments. c ELISA. BV-2 cells were treated with fowlerstefin (15 µg/ml) for 0–12 h. At various time points during the incubation, the culture supernatant was collected and the levels of TNF, IL-1α, IL-1β and IL-6 were analyzed by ELISA. Values are presented as the mean ± SD of three independent experiments. One-way ANOVA with Dunnett’s post-hoc test was performed as multiple comparisons with the control (NC) (for details, see Additional file 3: Text S1). ***P < 0.0001